Immortalized (Conditionally) Mouse Colonic Epithelial Cells (YAMC)

IGF2BP2 Recognizes and Binds to m6A Site of NCOA3 and Promotes mRNA Stability of NCOA3

Ulcerative colitis (UC), an idiopathic and chronic inflammatory disease, primarily targets the mucosal lining of the colon. This research endeavors to reveal the mechanism of insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) and nuclear receptor coactivator-3 (NCOA3) in UC-induced intestinal mucosal barrier dysfunction.

To assess the impact of IGF2BP2 on intestinal mucosal barrier integrity under DSS-induced colitis, they generated a murine model of UC by administering DSS to trigger colitis manifestations. Results showed that IGF2BP2 overexpression improved mucosal barrier dysfunction in these mice. In LPS-induced Caco-2 cells, NCOA3 expression was low, suggesting it might be a downstream target of IGF2BP2. RIP assays in YAMC cells showed NCOA3 mRNA was significantly enriched on IGF2BP2 compared to IgG controls, confirming their interaction (Fig. 1A). MeRIP assays indicated that increased IGF2BP2 in YAMC cells heightened the m6A modification of NCOA3 mRNA (Fig. 1D). mRNA stability tests revealed that IGF2BP2 upregulation enhanced NCOA3 mRNA levels and extended its half-life (Fig. 1E). DSS treatment reduced NCOA3 expression in mouse colons, but IGF2BP2 elevation restored it (Fig. 1F and G). Overall, IGF2BP2 binds to the m6A site of NCOA3, enhancing its mRNA stability via m6A modification.

Fig. 1. IGF2BP2 recognizes and binds to m6A site of NCOA3 and promotes mRNA stability of NCOA3 (Li R, Gu B, et al., 2025).

Pantothenate Protects the Intestinal Epithelial Barrier and Prevents Colitis in Adult Mice

HMOs (human milk oligosaccharides), particularly 2'-fucosyllactose (2'-FL), promote the growth of Bifidobacterium and Lactobacillus, which have been associated with several health benefits associated with breast milk. Schalich et al. determined how 2'-FL protects from intestinal inflammation in adulthood by altering the gut microbiota.

Through exploratory metabolomics, they identified pantothenate as a protective metabolite that fortifies the intestinal barrier against oxidative stress. They next studied how pantothenate could exert direct effects on intestinal epithelial cells to protect the intestinal barrier by using in vitro assays. They evaluated the effect of pantothenate on the oxidative stress-induced disruption of the epithelial barrier. H₂O₂ reduced the paracellular permeability in Caco₂ cells measured by transepithelial electrical resistance (TEER) in a time-dependent manner, which was significantly reduced by pantothenate treatment (Fig. 2A). Furthermore, H₂O₂-induced redistribution of ZO-1 from apical tight junctional complexes to the cytoplasmic compartment of YAMC cells was prevented by pantothenate (Fig. 2B). In order to determine how pantothenate reduces oxidative stress in YAMC cells they measured the activity of glutathione peroxidase (GPX), the enzyme with antioxidant properties that eliminates free radicals to reduce oxidative stress. Pantothenate at 0.1 mM increased GPX activity in YAMC cells compared to control cells (P = 0.0171, Fig. 2C).

Fig. 2. Pantothenate preserves the intestinal epithelial barrier and inhibits oxidative stress in intestinal epithelial cells (Schalich KM, Buendia MA, et al., 2024).